Kim, Hyun-Jin;Park, Young-Ha;Hwang, Seong-Su;Kim, Sang-Heum;Chung, Soo-Kyo
Nuclear Medicine and Molecular Imaging
/
v.40
no.1
/
pp.48-50
/
2006
A 38-year-old female with arthralgia in right elbow joint for 6 months was referred for a bone scan which showed diffuse uptakes of $^{99m}Tc$-MDP in the liver and spleen without hepatosplenomegaly. She had a history of hepatitis B vaccination 3 days ago. These uptakes were disappeared on the follow-up bone scan after 4 months. We suggest this transient diffuse hepatic uptake after vaccination of hepatitis B might be due to aluminum component within the hepatitis B vaccine as adjuvant.
PreS domain of Hepatitis B virus (HBV) surface antigen is a good candidate for an effective vaccine as it activates both B and T cells besides binding to hepatocytes. This report deals with overexpression and purification of adr subtype of surface antigen that is more prevalent in Pakistan. PreS region, comprising 119 aa preS1 region plus a 55 aa preS2 region plus 11 aa from the N-terminal S region, was inserted in pET21a+ vector, cloned in E. coli $DH5\alpha$ cells and expressed in E. coli BL21 codon+ cells. The conditions for over expression were optimized using different concentrations of IPTG (0.01-5 mM), and incubating the cells at different temperatures (23-$41^{\circ}C$) for different durations (0-6 h). The cells were grown under the given optimized conditions (0.5 mM IPTG concentration at $37^{\circ}C$ for 4 h), lysed by sonication and the protein was purified by ion exchange chromatography. On the average, 24.5 mg of recombinant protein was purified per liter of culture. The purified protein was later lyophilized and stored at $-80^{\circ}C$.
We examined the immunological properties of the recombinant hepatitis B surface antigen (r-HBsAg) which was expressed in mammalian cell (C127). The cross-immunity of r-HBsAg and plasma-derived hepatitis B surface antigen (p-HBsAg) were tested using Western blotting and ELISA with guinea pig polyclonal antibody and naturally infected human-derived antibody and the both antigens show the same results in their response pattern and intensity, which indicate they have a good cross-immunity. from the measurement of $ED_{50}$ after formalin- or heat-inactivation, both r-HBsAg and p-HBsAg and p-HBsAg showed $ED_{50}$ of 0.2-0.3 in formalin-inactivaton, while r-HBsAg was 0.05-0.09 and p-HBsAg was 0.03-0.07 in heat-inactivation, which means heat-inactivation method is 3-4 times superior in immunogenicity. In the immunopersistency test performed in guinea pig for the period of 3 months with two different adjuvants, antibody titer was 34.2 with muramyl dipeptide adjuvant, which was 1.8 times greater than the antibody titer of 18.9 with $AIPO_{4}$ adjuvant. the mutagenicity of r-HBsAg has the same cross-immunity with p-HBsAg, and heat-inactivation method and muramyl dipeptide adjuvant allow development of r-HBsAg vaccine with excellent immunogenicity.
Recombinant polioviruses have been developed by many research groups for use as vaccine vector because poliovirus induces mucosal immunity as well as humoral immunity through oral uptake. We assessed the potential use of poliovirus as a T-cell epitope carrier. Recombinant poliovirus V129 5L was constructed to have a substituted T-helper epitope from the core protein of Hepatitis B virus at neutralization antigenic site 1 on its VP1 capsid protein. The recombinant virus replicated less efficiently than type 1 poliovirus Mahoney strain. The V129 5L formed a little smaller plaques than the Mahoney strain and showed some 1.25 log unit lower titer at the peak in the one-step growth kinetics though it had similar growth profile to that of the Mahoney strain. Since V129 5L recombinant virus was genetically stable even after 24 successive passages in HeLa cells, the antigenic site 1 on VP1 capsid protein was confirmed for its ability of carrying T cell epitope. The genetic stability of V129 5L also indicated that recombinant poliovirus can be successfully utilized for the development of the multivalent vaccines.
The physicochmical properties of recombinant hepatitis B surface antigen (r-HBsAg), which was expressed in C127 mammalian cell were studied. Using roller bottle culture in DMEM supplemented with fetal bovine serum, 10-15 mg/L of r-HBsAg was produced with about 31% of purification yield. The purity of r-HBsAg by HPLC was 99.8% and electron microscopic examination showed homogeneous spherical particle with 22 nm in diameter, a morphological characteristic of HBsAg. The density of r-HBsAg by CsCI density gradient method was 1.19g/ml and the isoelectric point by Mono $P^{TM}$ HR 5/20 column was 4.6. The analysis of subunit protein pattern using SDS-PAGE followed by scanning densitometry gave 81.3% of S protein and 18.7% of pre-S protein. fluorophore-assisted-carbohydrate-electrophoresis analysis showed the relative amount of carbohydrate to protein was 1.7% and it smajr component was N-acetyl glucosamine, which was about 39% of total carbohydrate. The relative amount of lipid to protein determined by vanillin phosphoric acid method was 32.5% and its major component was phospholipid, which was about 70% of total lipid. The physicochemical properties of C127 mammalian cell-derved r-HBsAg are similar to those of p-HBsAg, suggesting that the r-HBsAg can be used in developing a new preventive vaccine against hepatitis B.
Chung, Eun Hee;Ma, Sang Hyuck;Hong, Young Jin;Kim, Kyung Hyo;Kim, Jong-Hyun;Lee, Jin A;Lee, Hoan Jong
Pediatric Infection and Vaccine
/
v.13
no.2
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pp.163-173
/
2006
Purpose : To reduce the number of injections necessary to vaccinate young infants, various combined vaccines have been developed. The $Comvax^{TM}$ manufactured by Merck & Co. is a combination of Hepatitis B and PRP-OMP conjugate Haemophilus influenzae Type b vaccine. The purpose of this study is to evaluate the immunogenicity and safety of $Comvax^{TM}$ in Korean infants. Methods : The infants who were vaccinated at 0 months of age with Hepatitis B vaccine, were recruited for this study after parental informed consent was obtained. The subjects were vaccinated with $Comvax^{TM}$ at 2 and 4 months of age. At each visit, infants were also immunized with DTaP, inactivated poliovirus vaccine, and pneumococcal vaccine when indicated. The serum anti-PRP and anti-HBs were measured at 2 months after the 1st dose(4 months age), and the 2nd dose(6 months age) by the ELISA and chemiluminescent microparticle immunoassay method, respectively. The local and systemic adverse reactions of vaccination were monitored for 3 consecutive days after each immunization. Results : Among sixty-five healthy infants(35 male infants) enrolled in this study; fifty eight(32 male infants) completed the scheduled immunizations. The geometric mean titers (GMTs) of anti-PRP at 2 months after the 1st dose and the 2nd dose were 1.96 ${\mu}g/mL$ (95% CI; 1.38~2.78) and 10.02 ${\mu}g/mL$ (95% CI; 7.04~14.26), respectively. Anti-PRP ${\geq}1.0$${\mu}g/mL$, was obtained in 63.2%(95% CI; 53.75~72.65) after 1 dose, and 96.6%(95% CI; 93.05~100) after 2 doses. The GMTs of anti-HBs were 38.32 mIU/mL(95% CI; 22.42~65.51), and 101.17 mIU/mL(95% CI; 65.94~155.25) at 2 month after the 1st dose and 2nd dose of $Comvax^{TM}$, respectively. Anti-HBs ${\geq}10$ mIU/mL was observed in 73.7%(95% CI; 65.07~82.33) after 1 dose and 94.8%(95% CI; 90.45~99.15) after 2 doses. Most of the adverse reactions after vaccination were mild. Irritability, the most common systemic reaction, was observed in 24.8%, followed by drowsiness(19.2%), poor feeding(19.2%) and fever(7.2%). Among the local reactions tenderness was observed in 25.6%, redness(${\geq}5$ mm) in 19.2% and swelling(${\geq}5$ mm) in 4.8%. Conclusion : The $Comvax^{TM}$ vaccine was highly immunogenic for PRP and safe in Korean infants. Although the hepatitis B vaccine component was administered at 0, 2, 4 months, this study showed good immunogenicity against HBsAg.
Purpose : This study was conducted to assess the current(2003-2005) prevalence of anti-HBs and immunologic memory for Hepatitis B vaccine in children from the central area of Korea. Methods : Subjects were chosen from children and adolescents who received tests for hepatitis B surface antigen(HBsAg) and anti-HBs at Dankook University Hospital from March 2003 to May 2005. Among these, antibodies to hepatitis B core antigen(IgG anti-HBc) were checked. A single booster vaccination was performed on children whose anti-HBs titers were under 10 mIU/mL. One month after booster vaccination we rechecked the anti-HBs titer. Results : A total of 3,277 subjects were tested for HBsAg/anti-HBs, and 1,913(58.4 percent) of them were positive for anti-HBs. Of these, 29 subjects(0.9 percent) were positive for HBsAg. Positive results for anti-HBs by age were 78.6 percent for 6-12 months of age, 62.7 percent for 1-3 years of age, 51.9 percent for 4-6 years of age, 49.5 percent for 7-12 years of age, 63.4 percent for 13-15 years of age and 72.2 percent for 16-18 years of age. The 80 subjects who were tested negative for HBsAg/anti-HBs received a single booster vaccine, 71 subjects were tested positive for antibodies. IgG anti-HBc titer was checked for 169 of the subjects, 5 subjects were positive. Conclusion : In our study, a significant anamnestic response was observed in 88.8 percent of children. This is believed to be a result of the relatively long immunologic memory effect of the hepatitis B vaccination in children from the central area of Korea.
Kim, Byung-Seok;Lim, Hyun-Sul;Lee, Kwan;Min, Young-Sun;Yoon, Young-Sil;Jeong, Hye-Sook
Journal of Preventive Medicine and Public Health
/
v.48
no.1
/
pp.53-61
/
2015
Objectives: The seroprevalence of hepatitis E virus (HEV) among high-risk groups overseas is high, but studies in these groups are rare in South Korea. We conducted the present study from April to November 2012 to obtain data on the seroprevalence and associated risk factors for HEV among slaughterhouse workers in South Korea. Methods: Slaughterhouse workers from 80 workplaces nationwide were surveyed in South Korea in 2012. The subjects comprised 1848 cases: 1434 slaughter workers and 414 residual products handlers. By visiting 80 slaughterhouses, which were mixed with 75 of which also performed residual products handling, we conducted a questionnaire survey for risk factors and obtained blood samples in order to determine the seropositivity and seroprevalence of HEV. Anti-HEV IgG and IgM were measured using HEV IgG and IgM enzyme-linked immunospecific assay kits and HEV antigen was measured by reverse transcription polymerase chain reaction (RT-PCR). Results: The seropositivity of anti-HEV IgG was 33.5% (slaughter workers 32.8% and residual products handlers 36.2%), and among the seropositive individuals the seroprevalence of anti-HEV IgM was 0.5% (slaughter workers 0.5%, residual products handlers 0.7%). The response rate of HEV-antigen as measured by RT-PCR was 0.2%. Risk factors significantly related to anti-HEV IgG seropositivity were age, sex, and working duration (slaughter workers only). Conclusions: There were significant risk factors (sex, age, and working duration) for HEV identified in our study. All three positive cases for HEV-antigen by RT-PCR were related to pig slaughter but without statistical significance. To prevent HEV, an educational program and working guidelines may be needed for high risk groups.
The Journal of the Korean life insurance medical association
/
v.3
no.1
/
pp.245-255
/
1986
To investigate the Positive rate of anti HBs and HBs Ag positive rate of the each group of vaccinated hepatitis B type and not-vaccinated, male 4150 female 2475, from May 1, 1984 to November 30, 1985 in the Medical department of Dae han Kyoyuk ins. Co., We examined into the HBsAg and Anti HBs by using the RPHA method and PHA method and the results which were adjusted statistically were as follows. 1. HBs Ag positive rate was 9.0%(228/2531)in 1984 and 7.0%(287/4069) in 1985, the positive rate in 1985 represented 2% lower than in 1984. 2. Anti HBs positive rate was 43.3%(1096/2531) in 1984 and 42.8%(1744/4069) in 1985, there was no significant change for two years. 3. Anti HBs positive rate in the group of the not-vaccinated hepatitis B type was 39.2%(869/2215) in 1984 and 38.8%(1333/3432)in 1985. 4. Anti HBs positive rate in the group of the vaccinated hepatitis B type more than once was 64.7%(419/647)in male and 71.5%(219/306) in female that was 66.9%(638/953) in total. 5. Anti HBs positive rate in the group of the vaccinated hepatitis B type three times was 68.7%(270/393) in male and 80.0%(156/195) in female and 72.4% in total. In this conclusion; The formation of Anti HBs in female showed more or less higher than male. 6. The cases which were detected HBs Ag and anti HBs at the same time were 4 in male and 3 in female and in the group of the vaccinated hepatitis B type, the number of HBs Ag positive cases were 8 in male and 10 in female. (On condition that we didn't distinguish $5{\mu}g/ml$, hepatitis B type vaccine, from $20{\mu}g/ml$ and after vaccinating, the lapsed time was not settled.)
Cha, Jihei;Kim, Han Wool;Lee, Soyoung;Cho, Hye Kyung;Ahn, Jong Gyun;Kim, Kyung-Hyo
Pediatric Infection and Vaccine
/
v.23
no.2
/
pp.117-127
/
2016
Purpose: Vaccine evaluation studies were initiated from 2000 by the Ministry of Food and Drug Safety to produce proper data about the safety and immunogenicity of vaccines. The purpose of this study was to review studies and reports on evaluation of vaccine such as immunogenicity, efficacy, effectiveness, safety and other related topics in order to find and analyze the data on the usefulness of each vaccine. Methods: From 2000 to 2014, the project "The vaccine evaluation" had been performed by several researchers, and studies and reports of vaccine evaluation. We reviewed the results and outcomes of studies regarding the evaluation of vaccine's usefulness and analyzed the possibilities of applying these data for establishing vaccine policies. For each vaccine, data analysis and organization were done according to evaluation fields. Results: A total of 83 studies were performed on vaccines from 2000 to 2014. For each vaccine, 8 studies were performed on BCG, 14 on DTaP/Td, 1 on poliovirus, 5 on Hib, 3 on pneumococcus, 11 on influenza, 3 on hepatitis A, 11 on MMR, 11 on varicella, and 16 on Japanese encephalitis. All studies were analyzed by the following evaluation area, such as safety, immunogenicity, seroprevalence, persistence of immunity, efficacy, effectiveness, vaccine evaluation methods, quality control product for vaccine, and others. Conclusions: Vaccine evaluation studies performed in Korea may be useful as references for establishing vaccination strategy and policy and could be used as baseline data for future studies on vaccine evaluation, vaccine policy establishment, and public/expert vaccine education in Korea.
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